Bioreactors have been used for cultivation of microbial organisms for production of various biological or chemical products in the pharmaceutical, beverage and biotechnological industry. A production bioreactor contains culture medium in a sterile environment that provides various nutrients required to support growth of the biological agents of interest. Conventional bioreactors use mechanically driven impellors to mix the liquid medium during cultivation. The bioreactors can be reused for the next batch of biological agents after cleaning and sterilization of the vessel. The procedure of cleaning and sterilization requires a significant amount of time and resources, especially, to monitor and to validate each cleaning step prior to reuse for production of biopharmaceutical products. Due to the high cost of construction, maintenance and operation of the conventional bioreactors, single use bioreactor systems made of disposable plastic material have become an attractive alternative.
While several mixing methods of liquid in disposable bioreactors have been proposed in recent years, none of them provides efficient mixing for large scale (greater than 1000 liters) without expensive operating machinery. For this reason, a number of non-invasive and/or disposable mixing systems that do not require an external mechanical operation have been developed. Many of these systems work well within certain size ranges, however, problems sometimes arise as larger mixing systems are attempted. Some relevant examples of prior art pneumatic mixing systems include the following.
U.S. Pat. No. 6,032,931, issued to Plunkett, discloses an aeration device for use in a pond. Compressed gas is supplied to a conduit to form bubbles as the gas/air exits from a series of apertures. As the bubbles rise, they drive turbines to rotate and thereby create additional mixing turbulence.
U.S. Pat. No. 6,322,056, issued to Drie describes a submarine type liquid mixer with aeration. The buoyancy shells provide a downwardly facing upwardly concave surface for capturing gas bubbles so as to provide a buoyancy force to the struts. The bubbles may be naturally involved within the liquid due to chemical processes or they may be released from a gas inflow into the tank. As the gases are captured by a series of shells, each in turn is displaced upwardly whereupon the gas is released at the top of tank. At this point, one of the shells loses its buoyancy while the lower shell has received gas bubbles, enabling it to be displaced and thus the motion of the shells is reversed. This up and down cyclic motion of the shells mixes the liquid in the tanks.
U.S. Pat. No. 6,406,624, issued to De Vos discloses a flocculation apparatus and apparatus for floating upwardly in a liquid and for moving downwardly in the liquid under the influence of gravity. The flocculation apparatus includes a paddle apparatus and a flotation and compressed gas discharge apparatus. A pressurized or compressed air line with a branch line extending upwardly into the flotation and compressed gas discharge apparatus is also provided. When gas is introduced through the lines into the gas discharge apparatus, the apparatus becomes increasingly buoyant and floats upwardly in the liquid within the basin and thereby moves the paddle apparatus and frame apparatus upwardly in the liquid as well. When the apparatus reaches the top, the compressed air is released and the frame apparatus along with the paddle apparatus are pulled downwardly in the liquid by gravity. During the upward and downward movement of the paddle members, the paddle members agitate or stir the liquid contained within the basin.
U.S. Pat. No. 6,390,455, issued to Lee et al. describes a bubble generating device having a float connected thereto. The object of the invention is to provide a bubble generating device that can be operated in a desired depth of water which ultimately is used to agitate the water and provide a supply of oxygen to the water. The device includes a porous portion which is connected to a source of air through a pipe to generate bubbles while the float maintains the apparatus at a desired level in a water container.
U.S. Pat. No. 5,645,346, issued to Thuna is directed to a food preparation blender with a rotating and vertically oscillating mixing blade. The blender includes a pressure plate which causes a first shaft to be raised, thus raising the mixing blades while mixing takes place.
U.S. Pat. No. 6,649,117, issued to Familletti discloses an improved reactor/fermentor apparatus useful for carrying out cell culture and fermentation. The apparatus utilizes novel design features to provide optimum agitation of the cells while minimizing mechanical shear force. The reactor is composed of two chambers; an upper, wider chamber and a lower, small diameter chamber which are connected by inwardly sloping side walls. Agitation is accomplished by utilizing a gently flowing centrally disposed gas stream.
U.S. Pat. No. 3,963,581, issued to Giacobbe et al. describes an air lift fermentor comprising in combination a hollow cylindrical body, vertically located and subdivided into three zones by a pair of diaphragms parallel to the axis of said cylindrical body, the central zone of which is destined to fermentation of the liquor, and the two lateral zones serve for recirculating the liquor itself, after its passage through a heat exchanger and an air distributor, both located near the bottom of said cylindrical body.
It is an objective of the present invention to provide a pneumatic bioreactor that is capable of efficiently and thoroughly mixing solutions without contamination. It is a further objective to such a reactor that can be scaled to relatively large sizes using the same technology. It is a still further objective of the invention to a bioreactor that can be produced in a disposable form. It is yet a further objective of the invention to provide a bioreactor that can be accurately controlled by internal pneumatic force, as to speed and mixing force applied to the solution without creating a foaming problem. Finally, it is an objective to provide a bioreactor that is simple and inexpensive to produce and to operate while fulfilling all of the described performance criteria.
While some of the objectives of the present invention are disclosed in the prior art, none of the inventions found include all of the requirements identified.